Monoclonal Antibody Development

Our services include:

• Design of antigens, immunization and screening strategies. This includes peptide design and decisions regarding the selection of multiple antigenic or branched peptides (MAPs) versus KLH (or other carrier protein)-conjugates with linear peptides, antigen purification and solubilization, and the optimization of screening protocols
• Conjugation of linear peptides to carrier molecules such as KLH or BSA. This includes quality control of peptide conjugates via SDS-PAGE gel analysis
• Hybridoma cell line production via traditional methods. This includes quality control of protein or peptide immunogen samples, initial screening of polyclonal antiserum via ELISA, Western blot, and in immune precipitation, fusion, post-fusion screening, positive well expansion and sub cloning of hybridomas secreting monoclonal antibodies with the desired specificity
• Optimization of existing hybridoma cell lines by selection cloning
• Production of monoclonal antibodies in species other than mice. Currently, we have rat (Y3-D10 from Lydia Sorkin, Karolinska) and rabbit (240E from Katherine Knight, Loyola) myeloma cell lines which have been tested and found capable of yielding antibody producing hybridoma cell lines. The resulting antibody secreting cell lines are useful in situations where investigators need monoclonal antibodies directed towards murine antigens, or when human antigens are not immunogenic in mice
• The facility offers services to wean hybridoma cell lines off selection media and into a variety of basal and serum free media for use in mini-bioreactors, immune precipitation or in vivo studies
• The cloning, expression and engineering of single chain antibody variable fragments (scFvs) or scFvs containing biological reporter molecules such as green fluorescent protein
• The selection of antigen specific antibodies and their fragments from phage display libraries. We have recently completed the construction of a high diversity phage display library containing genomic rat/mouse Ig sequences and have successfully selected antigen specific scFv bearing phage
• The facility also provides monoclonal antibodies directed to a number of biological tags and reported molecules. These include green fluorescent protein, glutathione S transferase, maltose binding protein, myc, 6-H, M13 phage (native or denatured), wheat germ agglutinin, E-tag expressed on scFv from the Hutchinson phage display library and other antibodies directed to members of the integrin and Ig-CAM families

Since each hybridoma development project is unique and successful project will often require that immunization and screening protocols be tailored to the unique needs of the investigator, we have the expertise available to assist investigators to design the exact screening mechanism to produce the desired reagent.

Contact resource staff for initial consultation and project scheduling (206) 667-3684.